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1、Evidence for NG2-glia Derived, Adult-Born FunctionalNeurons in the Hypothalamus,Sarah C. Robins1, Eric Trudel2, Olivia Rotondi1, Xiaohong Liu1, Tina Djogo1, Darya Kryzskaya1, Charles W. Bourque2, Maia V. Kokoeva1*,汇报人:学号:,1,神经胶质细胞文献回报,思路:,有文献报道,鼠科动物下丘脑有生成神经元的功能,但是是哪种细胞具有这种功能?(进行鉴定) 脑室膜细胞(ventricular
2、 tanycytes)被鉴定为是神经元生成的起源,但是研究发现,由此产生的神经元占总新神经元的很少部分,另一个起源是? 有报道下丘脑细胞能产生具有神经元标志物NeuN or HuC/D (Hu)的细胞,但是缺乏电生理的证明? Rivers et al用基因命运跟踪技术证明表达PDGFRa的细胞能在成年鼠的梨状皮层中产生投射神经元(但是其他人用Olig2-CreER or PDGFRa-CreER mice不能证明此结果或提出质疑), 那么NG2-glia是不是也可以产生neuron?,2,神经胶质细胞文献回报,这篇文章主要是运用转基因技术,失踪技术,对下丘脑中NG2-glia进行不同时期的观察
3、,并对最终分化而成的的带有神经元marker的细胞进行电生理鉴定,最终说明NG2-glia是成体下丘脑中新生神经元的主要来源.,3,神经胶质细胞文献回报,NG2-glia:,NG2-glia是以硫酸软骨素蛋白聚醣(NG2)和血小板衍生物生长因子受体alpha(PDGFRa)为标志物的神经胶质细胞; 占成体中枢神经系统细胞的10%; 能够分裂增殖,产生少突胶质细胞; 有文章报道,其在梨状皮层(piriform cortex, 和下丘脑一样都属于边缘系统)中能产生少量神经元,但是也有文章对此质疑;,4,神经胶质细胞文献回报,A为脑室灌注一次24h后,下丘脑实体成对出现BrdU+的细胞,说明由迁徙而
4、来的可能性不大;BC可以看出分裂细胞细胞广泛分布于下丘脑,没有富集现象;BD是七天灌注后立即取样观察和28天后观察的对比无明显分布,说明这些细胞的不是迁徙来的;E显示BrdU+ cell isNG2-glia,下丘脑中存在大量增值细胞,而非迁移过来的,大部分是NG2-glia,5,神经胶质细胞文献回报,有报道,Sox2+ cell能分化为神经元,那么NG2-glia如果能分化为神经元,也可能产生Sox2,A:Sox2+ cells主要分布在第三脑室附近和下丘脑实体;CDE:三种双阳性类型;ABF:37%+2.8%NG2+、Sox2+双标记,呈均匀分布,NG2是主要的增值细胞,6,神经胶质细胞文
5、献回报,Co-labelling with NG2 and Sox2 antibodies indicated that 37.0% +2.8 of hypothalamic NG2-glia robustly express Sox2 (Fig. 2A,B,F). Despite the fact that NG2-glia accounted for fewer than 10% of the total Sox2+ cell population, almost all (92.2%+2.8) of the hypothalamic BrdU+/Sox2+ cells also expr
6、essed NG2 (Fig. 2A,B,F), suggesting that NG2-glia are the only substantially proliferative subpopulation of Sox2+ cells in the adult hypothalamic parenchyma. three types of NG2-glia pairs according to Sox2 expression: i) substantial Sox2 expression in both cells; ii) no to barely detectable Sox2 exp
7、ression in both cells; or iii) Sox2 expression limited to one cell (Fig. 2CE), relate to differences in neural fates.,7,神经胶质细胞文献回报,BC:NG2是下丘脑主要的增值细胞;D-F:30days postTX 还有NG2细胞表达;L:7天和60天后,双阳性的绝度数量一样,2-7天少突胶质细胞迅速增多为重组得的保真性提供基础;GH:表面不表达星形胶质细胞的marker,即不分化为星形胶质细胞;LM:Tom+细胞中:NG2+数量和比率减少,而APC+相反,8,神经胶质细胞文献
8、回报,More than two thirds of the detected BrdU+/EdU+ cells expressed NG2 (Fig. 3B,C), indicating that adult hypothalamic NG2-glia can undergo multiple self-renewing divisions, in line with findings for the cerebral cortex 11; one of the primary functions of adult hypothalamic NG2-glia is to provide a
9、continuous supply of new oligodendrocytes;,9,神经胶质细胞文献回报,Figure S1 Fidelity of the NG2creER:tdTomato reporter. Mice were treated with TX twice daily for two days. (A) Cell types were determined by colocalisation of Tom with antibodies to NG2 to identify NG2-glia (arrowhead N), CD13 to identify pericy
10、tes (arrowhead P) and APC to identify oligodendrocytes (arrowhead O). (B) Quantification of results from A. Labelling with antibodies to HuC/D (C) and NeuN (D) confirmed that Tom+ neurons were not present at this timepoint.,重组实验的保真性研究:周皮细胞和已分化神经细胞不表达tom,10,神经胶质细胞文献回报,A-C:tom+和HuC/D+细胞被检出;C-D:a-tom+和
11、NeuN+被检出;G:双标记细胞随时间呈线性增长;H:tom+中Hu+和Neu+的比率不同,代表的时期不同,不同的偏好等原因,11,神经胶质细胞文献回报,A:第一次注射TX,后持续饮用含有BrdU的水直到,处死取样,发现Hu+/Tom+/BruU+的三标细胞;B: Hu+/Tom+中BruU+的比率和tom+中BruU+的比率没有明显不同 这组实验说明NG2-glia分化为了神经元,12,神经胶质细胞文献回报,B:对上一个实验进行补充:tom+ neurons表现出全或无的现象,自发的突出后电位(箭头所指)显示,这些tom+ neurons与周围神经元形成了回路。,C:tom+ neurons和tom- neurons在静息电位、电阻、电位和电容等特性的不同,可能是因为tom+还处于未成熟状态(齿状回非成熟颗粒细胞和成熟颗粒细胞类似),13,神经胶质细胞文献回报,谢谢!请指正交流,14,神经胶质细胞文献回报,